Methods and materials for treating lung disorders

ABSTRACT

This document provides methods and materials for treating lung conditions induced by inhalable insulin therapy. For example, compositions including a dried amnion tissue preparation and/or a dried stem cell preparation as well as methods for using such compositions to treat inhalable insulin induced lung conditions are provided.

CLAIM OF PRIORITY

The application is a Continuation of U.S. application Ser. No.14/984,738, which claims priority to U.S. Provisional Application Ser.No. 62/099,869 filed Jan. 5, 2015, U.S. Provisional Application Ser.No.62/166,233 filed May 26, 2015, and U.S. Provisional Application Ser.No. 62/240,076 filed Oct. 12, 2105, the entire contents of which arehereby incorporated by reference.

BACKGROUND 1. Technical Field

This document relates to methods and materials for treating lungdisorders (e.g., lung disorders induced by treatment with inhalableinsulin or exercise-induced pulmonary hemorrhage). For example, thisdocument provides methods and materials for using compositions (e.g.,inhalable formulations) that include a dried amnion tissue preparationand/or a dried stem cell preparation to treat lung disorders induced byinhalable insulin therapy. This document also provides methods andmaterials for using compositions (e.g., inhalable formulations) thatinclude a dried amnion tissue preparation and/or a dried stem cellpreparation in combination with an inhalable insulin therapy to reduceor prevent development of lung disorders induced by inhalable insulintherapy. This document also provides methods and materials for usingcompositions (e.g., inhalable formulations) that include a dried amniontissue preparation and/or a dried stem cell preparation to treat lungdisorders such as exercise-induced pulmonary hemorrhage.

2. Background Information

Inhalable insulin (Afrezza) is a rapidly acting form of insulin thatpatients with Type 1 or Type 2 diabetes can use to help with glycemiccontrol. Afreeza® can cause a decline in lung function over time asmeasured by forced expiratory volume (FEV₁) and common adverse reactionsinclude cough, and throat pain or irritation. Afreeza® also iscontraindicated in patients with chronic lung disease (e.g., asthma,chronic obstructive pulmonary disease (COPD), or other chronic lungdisease(s)). See, package insert for Afreeza®.

Exercise-induced pulmonary hemorrhage is a medical condition that refersto the presence of blood in lung airways in association with exercise.In some cases, between about 40 to 70 percent of horses may experienceblood in the trachea following a horse race.

SUMMARY

This document provides methods for using compositions (e.g., inhalableformulations) that include a dried amnion tissue preparation, a driedstem cell preparation, or both in combination, to treat lung disordersinduced by inhalable insulin therapy. Such compositions can beformulated for inhalation and used to treat any type of lung disorderinduced by inhalable insulin therapy. For example, the compositions canbe used to treat bronchospasms, asthma, COPD, chronic bronchitis,emphysema, pulmonary hypertension, asthma, interstitial lung disease,acute respiratory distress syndrome, pneumonia, lung infections, orpulmonary fibrosis that may be induced by inhalable insulin therapy.

This document also provides methods and materials for using compositions(e.g., inhalable formulations) that include a dried amnion tissuepreparation and/or a dried stem cell preparation in combination with aninhalable insulin therapy to reduce or prevent development of lungdisorders induced by inhalable insulin therapy. For example, a mammal(e.g., human) needing or desiring to take an inhalable insulin therapy(e.g., Afrezza®; MannKind Corp.) can be instructed to take a composition(e.g., inhalable formulations) that includes a dried amnion tissuepreparation, a dried stem cell preparation, or both to reduce or preventdevelopment of lung disorders induced by inhalable insulin therapy. Insome cases, a mammal (e.g., a human) with a lung disorder such as asthmaand chronic obstructive pulmonary disease who is warned not toadminister an inhalable insulin therapy because of that lung disordercan be treated with a composition (e.g., inhalable formulation) thatincludes a dried amnion tissue preparation and/or a dried stem cellpreparation either as an initial treatment or as a combination treatmenttogether with an inhalable insulin therapy. After the initial treatmentwith a composition (e.g., inhalable formulation) that includes a driedamnion tissue preparation and/or a dried stem cell preparation, themammal can be treated with an inhalable insulin therapy.

In another aspect, this document provides methods for using compositions(e.g., inhalable formulations) that include a dried amnion tissuepreparation, a dried stem cell preparation, or both in combination, totreat lung disorders such as exercise-induced pulmonary hemorrhage. Suchcompositions can be formulated for inhalation and used to treat any typeof exercise-induced pulmonary hemorrhage. For example, the compositionscan be used to treat pulmonary hemorrhage that may be induced racing orathletic activities. In some cases, a mammal (e.g., human, dog, orhorse) suffering from or suspected of suffering from exercise-inducedpulmonary hemorrhage can be instructed to take a composition (e.g.,inhalable formulations) that includes a dried amnion tissue preparation,a dried stem cell preparation, or both to reduce or prevent developmentof exercise-induced pulmonary hemorrhage.

In general, one aspect of this document features a method of treating amammal having a lung disorder induced by inhalable insulin therapy. Themethod comprises, or consists essentially of, administering, to themammal via inhalation, a composition comprising a dried amnion tissuepreparation lacking viable cells or a dried stem cell preparationlacking viable cells. The lung disorder can be asthma, chronicobstructive pulmonary disease, chronic bronchitis, emphysema, pulmonaryhypertension, interstitial lung disease, acute respiratory distresssyndrome, pneumonia, a lung infection, or pulmonary fibrosis. The lungdisorder can include bronchospasms. The composition can comprise thedried amnion tissue preparation and the dried stem cell preparation. Thecomposition can comprise a therapeutic agent, a growth factor, apolymer, a lipopolymer, a lung surfactant, or a pharmaceuticalexcipient. The composition can comprise the lung surfactant. The lungsurfactant can be dipalmitoylphosphatidyl-choline (DPPC). Thecomposition can comprise a pharmaceutical excipient. The composition cancomprise a growth factor. The method can comprise monitoring lungfunction of the mammal. The composition can comprise the dried amniontissue preparation. The dried amnion tissue preparation can comprise adried amnion tissue preparation prepared from about 1 mg to about 10 gof amnion tissue per kg of body weight of the mammal. The compositioncan comprise the dried stem cell preparation. The dried stem cellpreparation can comprise a dried stem cell preparation prepared fromabout 0.3 million to about 3 million stem cells per kg of body weight ofthe mammal. The dried amnion tissue preparation can be a human driedamnion tissue preparation. The dried stem cell preparation can be ahuman dried mesenchymal stem cell preparation.

In another aspect, this document features a method of treating a mammalneeding insulin in a manner that reduces the risk of developing a lungdisorder induced by inhalable insulin therapy. The method comprises, orconsists essentially of, (a) administering, to the mammal viainhalation, insulin, and (b) administering, to the mammal viainhalation, a composition comprising a dried amnion tissue preparationlacking viable cells or a dried stem cell preparation lacking viablecells, wherein administration of the composition reduces the developmentof a lung disorder induced by the administering of the insulin in step(a). The lung disorder can be asthma or chronic obstructive pulmonarydisease. The composition can comprise the dried amnion tissuepreparation and the dried stem cell preparation. The composition canfurther comprise a therapeutic agent, a growth factor, a polymer, alipopolymer, a lung surfactant, or a pharmaceutical excipient. Thecomposition can comprise the lung surfactant. The lung surfactant can beDPPC. The composition can comprise the pharmaceutical excipient. Thecomposition can comprise the growth factor. The method can furthercomprise monitoring lung function of the mammal. The composition cancomprise the dried amnion tissue preparation. The dried amnion tissuepreparation can comprise a dried amnion tissue preparation prepared fromabout 1 mg to about 10 g of amnion tissue per kg of body weight of themammal. The composition can comprise the dried stem cell preparation.The dried stem cell preparation can comprise a dried stem cellpreparation prepared from about 0.3 million to about 3 million stemcells per kg of body weight of the mammal. The dried amnion tissuepreparation can be a human dried amnion tissue preparation. The driedstem cell preparation can be a human dried mesenchymal stem cellpreparation.

In another aspect, this document features a method of treating a mammalneeding insulin and having a lung disorder limiting the use of inhalableinsulin. The method comprises, or consists essentially of, (a)administering, to the mammal via inhalation, a composition comprising adried amnion tissue preparation lacking viable cells or a dried stemcell preparation lacking viable cells, wherein administration of thecomposition reduces the symptoms of the lung disorder, and (b)administering, to the mammal via inhalation, insulin. The lung disordercan be asthma or chronic obstructive pulmonary disease. The compositioncan comprise the dried amnion tissue preparation and the dried stem cellpreparation. The composition can further comprise a therapeutic agent, agrowth factor, a polymer, a lipopolymer, a lung surfactant, or apharmaceutical excipient. The method can further comprise monitoringlung function of the mammal. The composition can comprise the driedamnion tissue preparation. The dried amnion tissue preparation cancomprise a dried amnion tissue preparation prepared from about 1 mg toabout 10 g of amnion tissue per kg of body weight of the mammal. Thecomposition can comprise the dried stem cell preparation. The dried stemcell preparation can comprise a dried stem cell preparation preparedfrom about 0.3 million to about 3 million stem cells per kg of bodyweight of the mammal. The dried amnion tissue preparation can be a humandried amnion tissue preparation. The dried stem cell preparation can bea human dried mesenchymal stem cell preparation.

In another aspect, this document features a method of treating a mammalhaving exercise-induced pulmonary hemorrhage. The method comprises, orconsists essentially of, administering, to the mammal via inhalation, acomposition comprising a dried amnion tissue preparation lacking viablecells or a dried stem cell preparation lacking viable cells. The mammalcan be a horse. The mammal can be a racing horse. The composition cancomprise the dried amnion tissue preparation and the dried stem cellpreparation. The composition can further comprise a therapeutic agent, agrowth factor, a polymer, a lipopolymer, a lung surfactant, or apharmaceutical excipient. The composition can comprise the lungsurfactant. The lung surfactant can be dipalmitoylphosphatidyl-choline(DPPC). The composition can comprise the pharmaceutical excipient. Thecomposition can comprise the growth factor. The method can furthercomprise monitoring a lung of the mammal for bleeding. The compositioncan comprise the dried amnion tissue preparation. The dried amniontissue preparation can comprise a dried amnion tissue preparationprepared from about 1 mg to about 10 g of amnion tissue per kg of bodyweight of the mammal. The composition can comprise the dried stem cellpreparation. The dried stem cell preparation can comprise a dried stemcell preparation prepared from about 0.3 million to about 3 million stemcells per kg of body weight of the mammal. The dried amnion tissuepreparation can be a horse dried amnion tissue preparation. The driedstem cell preparation can be a horse dried mesenchymal stem cellpreparation.

Unless otherwise defined, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention pertains. Although methods and materialssimilar or equivalent to those described herein can be used to practicethe invention, suitable methods and materials are described below. Allpublications, patent applications, patents, and other referencesmentioned herein are incorporated by reference in their entirety. Incase of conflict, the present specification, including definitions, willcontrol. In addition, the materials, methods, and examples areillustrative only and not intended to be limiting.

The details of one or more embodiments of the invention are set forth inthe accompanying drawings and the description below. Other features,objects, and advantages of the invention will be apparent from thedescription and drawings, and from the claims. The word “comprising” inthe claims may be replaced by “consisting essentially of” or with“consisting of,” according to standard practice in patent law.

DETAILED DESCRIPTION

In general, this document provides methods and materials for treatinglung disorders and conditions induced by inhalable insulin (e.g.,Afrezza®) therapy using compositions that include a dried amnion tissuepreparation (e.g., dried human amnion tissue preparation) and/or a driedstem cell preparation. Such lung disorders and conditions can includebronchospasms, COPD, chronic bronchitis, asthma, emphysema, pulmonaryhypertension, interstitial lung disease, pulmonary fibrosis, pneumonia,lung infections, or acute respiratory distress syndrome.

This document also provides methods and materials for using compositions(e.g., inhalable formulations) that include a dried amnion tissuepreparation and/or a dried stem cell preparation in combination with aninhalable insulin therapy to reduce or prevent development of lungdisorders induced by inhalable insulin therapy. In some cases, a mammal(e.g., a human) with a lung disorder such as asthma and chronicobstructive pulmonary disease who is warned not to administer aninhalable insulin therapy because of that lung disorder can be treatedwith a composition (e.g., inhalable formulation) that includes a driedamnion tissue preparation and/or a dried stem cell preparation either asan initial treatment or as a combination treatment together with aninhalable insulin therapy. After the initial treatment with acomposition (e.g., inhalable formulation) that includes a dried amniontissue preparation and/or a dried stem cell preparation, the mammal canbe treated with an inhalable insulin therapy.

In some cases, a composition (e.g., an inhalable formulation) thatincludes a dried amnion tissue preparation and/or a dried stem cellpreparation can be administered to humans who smoke tobacco products(e.g., cigarettes, cigars, or pipes) or to humans with a history ofsmoking tobacco products (e.g., cigarettes, cigars, or pipes) to reducethe severity of symptoms (e.g., lung symptoms) of smoking or to reducethe development of symptoms (e.g., lung symptoms) of smoking. Forexample, a human who smokes cigarettes can be administered a composition(e.g., an inhalable formulation) that includes a dried amnion tissuepreparation and/or a dried stem cell preparation to reduce the severityof a chronic smoker's cough, a gravelly voice, and/or shortness ofbreath.

In another aspect, this document provides methods and materials fortreating lung disorders and conditions such as exercise-inducedpulmonary hemorrhage using compositions that include a dried amniontissue preparation (e.g., dried human amnion tissue preparation) and/ora dried stem cell preparation.

In some cases, a composition (e.g., an inhalable formulation) thatincludes a dried amnion tissue preparation and/or a dried stem cellpreparation can be administered to a mammal suffering from or suspectedto suffer from exercise-induced pulmonary hemorrhage. Examples of suchmammals include, without limitation, horses (e.g., race horses), dogs(e.g., racing dogs), or humans (e.g., human athletes).

The term “dried amnion tissue preparation” as used herein refers to apreparation of amnion tissue or amnion material that is dried to have awater content that is less than about 8 percent (e.g., less than about 7percent, less than about 6 percent, less than about 5 percent, less thanabout 4 percent, less than about 3 percent, less than about 2 percent,or less than about 1 percent). The term “dried stem cell preparation” asused herein refers to a preparation of stem cells or stem cell materialthat is dried to have a water content that is less than about 8 percent(e.g., less than about 7 percent, less than about 6 percent, less thanabout 5 percent, less than about 4 percent, less than about 3 percent,less than about 2 percent, or less than about 1 percent). In some cases,a dried amnion tissue preparation or a dried stem cell preparation canhave a water content that is between about 0.1 percent and about 8percent (e.g., between about 0.5 percent and about 8 percent, betweenabout 1 percent and about 8 percent, between about 0.1 percent and about5 percent, between about 0.1 percent and about 4 percent, between about0.1 percent and about 3 percent, between about 0.5 percent and about 5percent, or between about 1 percent and about 4 percent). An amniontissue preparation or stem cell preparation can be dried using anyappropriate technique such as micronization, vacuum drying, spraydrying, freeze drying, or combinations thereof In some cases, an amniontissue preparation or stem cell preparation can be dried as describedelsewhere (e.g., U.S. Pat. No. 5,656,498).

A dried amnion tissue preparation can contain viable cells, non-viablecells, or a combination thereof. For example, a dried amnion tissuepreparation can be a preparation of amnion tissue or amnion materialwhere all the cells were removed, killed, or lysed such that the driedamnion tissue preparation lacks viable cells. In some cases, a driedamnion tissue preparation can be a preparation of amnion tissue oramnion material that was exposed to one or more physical and/or chemicaltreatments that killed, fixed, or lysed the cells of the amnion tissueor amnion material such that the dried amnion tissue preparation lacksviable cells. For example, temperature (e.g., rapid freezing or rapidfreezing-thawing), force and pressure, and/or electrical disruption canbe used to kill or lyse cells within amnion tissue or amnion material toproduce a dried amnion tissue preparation that lacks viable cells.

In some cases, amnion tissue or amnion material can be obtained and thentreated in a manner designed to lyse all the cells within the amniontissue or amnion material. In these cases, the resulting material (e.g.,matrix material and cellular remnants from lysed cells) can be dried toform a dried amnion tissue preparation that lacks viable cells.

In some cases, a dried amnion preparation can be prepared from humanamnion tissue. For example, human amnion tissue can be harvested,processed to remove, kill, or lyse cells or to remove blood, and driedto form a dried amnion tissue preparation. In some cases, human amniontissue can be processed to remove blood prior to forming a dried amniontissue preparation. In some cases, human amnion tissue can be processedwithout removing cells or blood prior to forming a dried amnion tissuepreparation.

An example of a dried amnion tissue preparation includes, withoutlimitation, a dried human amnion tissue preparation that lacks viablecells. In some cases, a dried amnion tissue preparation can be obtainedfrom MiMedX® or a tissue bank (e.g., a human tissue bank).

In some cases, an amnion tissue preparation containing viable cells canbe used in place of or in addition to a dried amnion tissue preparationlacking viable cells to treat a lung disorder or condition such asexercise-induced pulmonary hemorrhage.

A dried stem cell preparation can contain viable stem cells, non-viablestem cells, or a combination thereof. For example, a dried stem cellpreparation can be a preparation of stem cell or stem cell materialwhere all the stem cells were killed, fixed, or lysed such that thedried stem cell preparation lacks viable stem cells. In some cases, adried stem cell preparation can be a preparation of stem cells or stemcell material that was exposed to one or more physical and/or chemicaltreatments that killed, fixed, or lysed the stem cells such that thedried stem cell preparation lacks viable stem cells. For example,temperature (e.g., rapid freezing or rapid freezing-thawing), force andpressure, and/or electrical disruption can be used to kill or lyse stemcells to produce a dried stem cell preparation that lacks viable stemcells.

In some cases, a stem cell culture can be obtained and then treated in amanner designed to lyse all the stem cells. In these cases, theresulting material (e.g., cellular remnants from lysed stem cells) canbe dried to form a dried stem cell preparation that lacks viable stemcells.

Examples of dried stem cell preparations include, without limitation, adried lung stem cell preparation such as a lung epithelial progenitorcell preparation, a dried mesenchymal stem cell (MSC) preparation (e.g.,a MSC preparation obtained from fat tissue or bone marrow), a driedumbilical cord blood stem cell preparation, a dried embryonic stem cellpreparation, and a dried human induced pluripotent stem cellpreparation.

In some cases, dried stem cell preparations are prepared from culturesof stem cells. For example, a culture containing from about 25 millionto about 25 billion stem cells can be used to make a dried stem cellpreparation. In some cases, from about 0.3 million to about 3 million(e.g., from about 0.3 million to about 3 million, from about 0.5 millionto about 3 million, from about 0.75 million to about 3 million, fromabout 1 million to about 3 million, from about 1.5 million to about 3million, from about 0.3 million to about 2.5 million, from about 0.3million to about 2.0 million, from about 0.3 million to about 1.5million, from about 0.3 million to about 1.0 million, from about 0.5million to about 2.5 million, from about 0.75 million to about 2.0million, from about 0.8 million to about 1.5 million) stem cells per kgof body weight of a mammal (e.g., a human) to be treated can be used tomake a dried stem cell preparation for administration to that mammal. Insome cases, a dried stem cell preparation can be obtained commerciallyfrom Stemedica Cell Technologies, Inc.

In some cases, a dried stem cell preparation can be prepared by washinga culture of stem cells in saline (e.g., phosphate buffered saline) toremove culture medium, evaporating to remove wash medium, adding asolution (e.g., saline, water, or a water and sugar solution) to theresulting stem cell preparation, and repeating the evaporation step.After the second evaporation step, the stem cell preparation can beformulated into a powder that can be used as a dried stem cellpreparation.

In some cases, a stem cell tissue preparation containing viable cellscan be used in place of or in addition to a dried stem cell preparationlacking viable cells to treat a lung disorder or condition such asexercise-induced pulmonary hemorrhage.

Typically, a composition described herein (e.g., a compositioncontaining a dried amnion tissue preparation, a dried stem cellpreparation, or both a dried amnion tissue preparation and a dried stemcell preparation) is administered via inhalation, and the dried amniontissue preparation and/or dried stem cell preparation will have aparticle size suitable for delivery to the respiratory tract of a user(e.g., a mammal such as a human, dog, cat, horse, cow, pig, sheep, goat,or monkey). For example, the dried amnion tissue preparation and/ordried stem cell preparation can have a particle size ranging from about0.1 μm to about 25 μm (e.g., from about 0.5 μm to about 25 from about0.75 μm to about 25 from about 1μm to about 25 from about 0.1 μm toabout 15 from about 0.1 μm to about 10 from about 0.1 μm to about 7.5from about 0.1 μm to about 5 from about 0.75 μm to about 7.5 or fromabout 1 μm to about 5 μm).

In some cases, a dried amnion tissue preparation and/or a dried stemcell preparation can have a particle size suitable for delivery to theupper respiratory tract (URT). The URT includes the nose, sinuses,pharynx and larynx. For example, a dried amnion tissue preparationand/or a dried stem cell preparation having a particle size ranging fromabout 5 μm to about 25 from about 5 μm to about 15 or from about 5 μm toabout 10 μm can be used to treat a disorder of the URT.

In some cases, a dried amnion tissue preparation and/or a dried stemcell preparation can have a particle size suitable for delivery to thelower respiratory tract (LRT), which includes the trachea, upperbronchi, and lungs, and be used to treat a lung disorder such asexercise-induced pulmonary hemorrhage. For example, a dried amniontissue preparation and/or a dried stem cell preparation having aparticle size ranging from about 0.1 μm to about 5 (e.g., from about 0.5μm to about 5 from about 0.75 to about 5 from about 1μm to about 5 fromabout 0.1 μm to about 2 from about 0.1 μm to about 1 from about 0.1 μmto about 0.75 μm) can be used to treat a lung disorder such asexercise-induced pulmonary hemorrhage.

In some cases, a composition that includes a dried amnion tissuepreparation and/or a dried stem cell preparation also can include one ormore therapeutic agents, one or more anti-inflammatory agents (e.g.,non-steroidal anti-inflammatory drugs, dexamethasone or other type ofglucocorticoid steroids), one or more growth factors (e.g., plateletderived growth factor PDGF, epithelial growth factor (EGF), fibroblastgrowth factor-2 (FGF2), or stem cell factor (SCF)), one or more lungsurfactants (e.g., DPPC), and/or one or more antimicrobial agents (e.g.,antibiotics such as kanamycin, neomycin, streptomycin, or gentamicin oran antifungal agent).

Compositions containing a dried amnion tissue preparation and/or a driedstem cell preparation can be formulated as inhalable compositions usingone or more pharmaceutically acceptable carriers or excipients. In somecases, a dried amnion tissue preparation and/or a dried stem cellpreparation can be formulated into microparticles that contain solidlipid nanoparticles. In some cases, the particles of a dried amniontissue preparation and/or a dried stem cell preparation can be coated orencapsulated for delivery to an airway (e.g., a lung) via an aerosolbased inhaler or a dry powder inhaler.

As described herein, lung disorders can be treated by administering(e.g., via inhalation) an effective amount of a composition thatincludes a dried amnion tissue preparation described herein and/or adried stem cell preparation described herein. Effective amounts ofcompositions described herein can be determined by a physician, takinginto account various factors such as overall health status, body weight,sex, diet, time and route of administration, other medications, and anyother relevant clinical factors. As used herein, an “effective amount”or “therapeutically effective amount” of a composition is the amountthat is sufficient to provide a beneficial effect to the subject towhich the composition or preparations are delivered. The effectiveamount can be the amount effective to achieve an improved survival rate,a more rapid recovery, an improvement in the quality of life, or animprovement or elimination of one or more symptoms associated with asubject's lung disorder (e.g., exercise-induced pulmonary hemorrhage).

In some embodiments, the methods include delivering, to the subject, adried amnion tissue preparation made with from about 0.01 mg to about 10g (e.g., from about 0.01 mg to about 10 g, from about 0.1 mg to about 10g, from about 1 mg to about 10 g, from about 10 mg to about 10 g, fromabout 100 mg to about 10 g, from about 1 g to about 10 g, from about0.01 mg to about 5 g, from about 0.01 mg to about 1 g, from about 0.01mg to about 100 mg, from about 10 mg to about 5 g, from about 100 mg toabout 1 g, or from about 1 g to about 5 g) of amnion tissue per kg bodyweight of the subject being treated.

In some embodiments, the methods include delivering, to the subject, adried stem cell preparation made from about 0.3 million to about 3million (e.g., from about 0.5 million to about 3 million, from about0.75 million to about 3 million, from about 1 million to about 3million, from about 1.5 million to about 3 million, from about 0.3million to about 2.5 million, from about 0.3 million to about 2.0million, from about 0.3 million to about 1.5 million, from about 0.3million to about 1.0 million, from about 0.5 million to about 2.5million, from about 0.75 million to about 2.0 million, from about 0.8million to about 1.5 million) stem cells per kg body weight of thesubject being treated.

In some embodiments, the compositions containing a dried amnion tissuepreparation described herein and/or a dried stem cell preparationdescribed herein are delivered to the subject by inhalation only once.In some embodiments, multiple (e.g., two, three, four, five, six, seven,eight, nine, 10, 11, 12, 13, 14, 15, or 20 or more) deliveries are madeby inhalation. For example, multiple deliveries of a dried amnion tissuepreparation described herein and/or a dried stem cell preparationdescribed herein can be made over the course of several (e.g., two,three, four, five, six, seven, eight, nine, 10, 14, 21, 28, or 31 ormore) consecutive days (e.g., one delivery each day for seven days orone delivery every other day for seven days). In some cases, a driedamnion tissue preparation described herein and/or a dried stem cellpreparation described herein can be delivered from about two to fourtimes a day to about once per month. In some cases, a dried amniontissue preparation described herein and/or a dried stem cell preparationdescribed herein can be delivered to a subject for several months (e.g.,one delivery per month for six months, or one delivery per week for twomonths).

A dried amnion tissue preparation described herein and/or a dried stemcell preparation described herein can be delivered to a subject atvarious time points after diagnosis with a lung disorder (e.g.,exercise-induced pulmonary hemorrhage). For example, a dried amniontissue preparation described herein and/or a dried stem cell preparationdescribed herein can be delivered immediately following diagnosis of alung disorder (e.g., exercise-induced pulmonary hemorrhage). In somecases, a dried amnion tissue preparation described herein and/or a driedstem cell preparation described herein can be delivered to a subjectless than 10 (e.g., 9, 8, 7, 6, 5, 4, 3, 2, or 1) days after diagnosiswith a lung disorder (e.g., exercise-induced pulmonary hemorrhage).

The subject can be any mammal, e.g., a human (e.g., a human patient) ora non-human primate (e.g., chimpanzee, baboon, or monkey), a mouse, arat, a rabbit, a guinea pig, a gerbil, a hamster, a horse, a type oflivestock (e.g., cow, pig, sheep, or goat), a dog, or a cat. In somecases, the subject can be any appropriate mammal such as a human (e.g.,a human athlete), a horse (e.g., a racing horse), or a dog (e.g., aracing dog) when treating exercise-induced pulmonary hemorrhage.

A composition described herein can be administered to a subject as acombination therapy with another treatment used to treat a lungdisorder. For example, the combination therapy can include administeringto the subject (e.g., a human patient) one or more additional agentsthat provide a therapeutic benefit to the subject who has, or is at riskof developing a lung disorder. In some cases, the composition and theone or more additional agents can be administered at the same time. Insome cases, the composition can be administered first, and the one ormore additional agents administered second, or vice versa.

The efficacy of a given treatment in treating a particular lung disordercan be defined as an improvement of one or more symptoms of the lungdisorder by at least 5% (e.g., at least 10%, at least 15%, at least 20%,at least 25%, at least 30%, at least 40%, at least 50%, at least 55%, atleast 60%, at least 65% or more). In some cases, efficacy of a treatmentwith a composition containing a dried amnion tissue preparation and/or adried stem cell preparation can be determined from the stabilization ofone or more symptoms associated with the lung disorder (i.e., thetreatments curtail the worsening of one or more symptoms of the lungdisorder).

In some cases, the methods described herein can include monitoring thelung disorder in the subject to, for example, determine if the disorderis improving with treatment. Any appropriate method can be used tomonitor a lung disorder. For example, for COPD patients, lung function(e.g., using a spirometer or arterial blood gas test) can be monitored.For exercise-induced pulmonary hemorrhage patients, clinical techniquesdesigned to detect the presence of blood in lung airways can be used.

Compositions that include a dried amnion tissue preparation describedherein and/or a dried stem cell preparation described herein can becombined with packaging material and sold as a kit. The packagingmaterial included in a kit typically contains instructions or a labeldescribing how the composition can be administered via inhalation. A kitalso can include an inhaler such as a unit dose inhaler or a multipledose inhaler. The term “unit dose inhaler” refers to an inhaler thatdelivers a single dose of a dry powder formulation by inhalation to auser. Typically, a unit dose inhaler contains a single container thatholds or contains an inhalable formulation. It will be appreciated thatin some cases, multiple unit doses are required to provide a user with aspecified dosage. The term “multiple dose inhaler” refers to an inhalerhaving two or more containers, each container comprising a pre-metereddose of a dry powder medicament, and the inhaler delivers a single doseof a medicament powder by inhalation at any one time.

As used herein a “unit dose” refers to a pre-metered formulation (e.g.,a dry powder formulation) for inhalation. In some cases, a unit dose canbe a single container having multiple doses of formulation that can bedelivered by inhalation as metered single amounts. A unit dosecartridge/container can contain a single dose. In some cases, it caninclude multiple individually accessible compartments, each containing aunit dose.

OTHER EMBODIMENTS

It is to be understood that while the invention has been described inconjunction with the detailed description thereof, the foregoingdescription is intended to illustrate and not limit the scope of theinvention, which is defined by the scope of the appended claims. Otheraspects, advantages, and modifications are within the scope of thefollowing claims.

What is claimed is:
 1. A composition comprising: a dried amnion tissuepreparation lacking viable cells, wherein the composition is aninhalable formulation.
 2. The composition of claim 1, furthercomprising: a dried stem cell preparation lacking viable cells, whereinsaid dried stem cell preparation is a dried umbilical cord-, fattissue-, or bone marrow-derived stem cell preparation of cellularremnants from lysed stem cells.
 3. The composition of claim 2, whereinsaid dried stem cell preparation is a dried umbilical cord-derived stemcell preparation.
 4. The composition of claim 2, wherein said dried stemcell preparation and said dried amnion tissue preparation are in theform of particles ranging in size from about 0.1 μm to about 10 μm. 5.The composition of claim 2, wherein said dried stem cell preparation andsaid dried amnion tissue preparation are in the form of particlesranging in size from about 1 μm to about 5 μm.
 6. The composition ofclaim 1, wherein said composition comprises from about 5 mg and about 5g of said dried amnion tissue preparation.
 7. The composition of claim1, wherein said composition comprises from about 10 mg and about 1 g ofsaid dried amnion tissue preparation.
 8. The composition of claim 4,wherein the particles are coated or encapsulated.
 9. The composition ofclaim 1, further comprising a therapeutic agent, a growth factor, apolymer, a lipopolymer, a lung surfactant, or a pharmaceuticalexcipient.
 10. An inhaler selected from a metered dose inhaler and a drypowder inhaler, the inhaler comprising an inhalable formulationcomprising: a dried amnion tissue preparation lacking viable cells. 11.The inhaler of claim 10, wherein the dried amnion tissue preparation isin the form of particles ranging in size from about 0.1 μm to about 10μm.
 12. The inhaler of claim 10, wherein the dried amnion tissuepreparation is in the form of particles ranging in size from about 1 μmto about 5 μm.
 13. The inhaler of claim 10, wherein the inhalableformulation comprises from about 5 mg and about 5 g of said dried amniontissue preparation.
 14. The inhaler of claim 10, wherein the inhalableformulation comprises from about 10 mg and about 1 g of said driedamnion tissue preparation.
 15. The inhaler of claim 11, wherein theparticles are coated or encapsulated.
 16. The inhaler of claim 10,further comprising a therapeutic agent, a growth factor, a polymer, alipopolymer, a lung surfactant, or a pharmaceutical excipient.
 17. Amethod comprising: administering to a lung of a subject an inhalableformulation comprising a dried amnion tissue preparation lacking viablecells.
 18. The method of claim 17, wherein the inhalable formulationfurther comprises a dried stem cell preparation lacking viable cells,wherein said dried stem cell preparation is a dried umbilical cord-, fattissue-, or bone marrow-derived stem cell preparation of cellularremnants from lysed stem cells.